CN116419753A

CN116419753A - Heterocyclic compounds as BCL-2 inhibitors

Info

Publication number: CN116419753A
Application number: CN202180057392.3A
Authority: CN
Inventors: 孔祥龙; 周超; 郑之祥
Original assignee: Beijing Innocare Pharma Tech Co Ltd
Current assignee: Beijing Innocare Pharma Tech Co Ltd
Priority date: 2020-08-06
Filing date: 2021-08-02
Publication date: 2023-07-11
Also published as: CA3190686A1; JP2023537032A; WO2022028353A1; AU2021319597A1; MX2023001618A; US20230271959A1; KR20230065986A; CN114057728A; EP4194454A1

Abstract

The present invention relates to compounds, pharmaceutical compositions containing them, and methods for their preparation, and their use as BCL-2 inhibitors. The compound is a compound shown in a formula (I), or an isomer, a prodrug, a solvate, a stable isotope derivative or a pharmaceutically acceptable salt thereof. The invention also relates to the use of said compounds for the treatment or prophylaxis of BCL-2 mediated diseases such as tumors and methods of using the same for the treatment of said diseases.

Description

Heterocyclic compounds as BCL-2 inhibitors

Technical Field

The present invention relates to compounds, pharmaceutical compositions containing them and their use as B-cell lymphoma-2 (BCL-2) inhibitors. More particularly, the present invention provides novel compounds which are BCL-2 inhibitors, pharmaceutical compositions containing such compounds and the use of the compounds to treat or prevent BCL-2 mediated related diseases and disorders, such as tumors. The invention also relates to a method for preparing said compounds.

Background

The BCL-2 family of proteins is one of the core regulatory mechanisms of apoptosis (also known as programmed cell death) that can receive and transmit intrinsic intracellular signals or external environmental stress signals, such as nutritional or hypoxic stress, DNA damage, oncogene overactivation, endoplasmic reticulum stress, etc., mainly playing a leading role in the intrinsic apoptosis pathway (Intrinsic pathway). The BCL-2 (B-cell lymphoma-2) protein was first discovered in 1986 and expressed from the BCL-2 gene. The BCL-2 gene is a protooncogene, and its expressed protein is called BCL-2 family protein. The total 27 proteins of BCL-2 family in human body can be divided into 3 subclasses according to function and sequence analysis, and the first subclass is antagonistic to apoptosis and comprises BCL-XL, BCL-2, BCL-W, MCL-1 and BFL-1, which are mainly positioned on mitochondria to protect the mitochondria from adverse injury; the other two subclasses are apoptosis-promoting, one is the end effector of mitochondrial injury, including BAX and BAK. The rest is classified into BH3 subclass, and can directly induce various cell adversity stress signals. Antagonizing and promoting the dynamic balance of interactions between apoptotic proteins to determine the death fate of cells. BCL-2 proteins that antagonize apoptosis are closely related to tumors, with about 50% of tumors (e.g., leukemia, rectal cancer, prostate cancer, etc.) present with aberrant overexpression of BCL-2 family proteins, wherein aberrant BCL-2 activity is prevalent in hematological tumors. Multiple signaling pathways, such as JAK-STAT, NFkB, UPP (ubiquitin-proteome), can cause overexpression of BCL-2 protein that antagonizes apoptosis.

The high expression of BCL-2 family antagonistic apoptosis proteins is related to drug resistance of various tumors, for example, the overexpression of BCL-2 antagonistic apoptosis proteins can enable tumor cells to evade apoptosis caused by anti-tumor drugs, so that drug resistance is caused. It has been shown that inhibition of BCL-2 family proteins can inhibit tumor neovascularization and thus tumor metastasis (Benjamin, d.; isaac, j.et al j.clin.oncol.2008, 26 (25), 4180). Thus, targeted inhibition of the BCL-2 family anti-apoptotic proteins can inhibit tumor development, progression and resistance.

Although there have been 20 reports of more than 20 small molecule inhibitors targeting the BCL-2 family, few have entered clinical trials, obatocrax in terraced tiles has only obtained partial responses in 1 out of 26 Chronic Lymphocytic Leukemia (CLL) treated, and has a strong neurotoxicity, ending the development in 2013; navitocrax (ABT-263) developed by Abovir, while exhibiting 50% good response rate in phase I dose ramp-up experiments in patients with recurrent or refractory lymphoid malignancies, also exhibited very strong targeted toxicity of BCL-XL: such as thrombocytopenia and severe anemia. Venetoclax (ABT-199), which is developed by combining Abbe's and Roche, is a highly selective BCL-2 inhibitor (Andrew, J.; joel, D.et al Nature Medicine,2013, 19 (2), 202), and has been greatly improved in Objective Remission Rate (ORR) and complete remission rate (CR) by combined administration with Ibutinib and the like in the treatment of recurrent/refractory Chronic Lymphocytic Leukemia (CLL), mantle Cell Lymphoma (MCL), multiple Myeloma (MM) and the like, but still has toxic and side effects such as leukopenia, anemia, diarrhea, dizziness, fatigue, susceptibility to infection, and the like, and serious toxic and side effects including pneumonia, anemia, hyperpyrexia and the like. There is therefore a need to develop BCL-2 selective inhibitors with high activity and low toxic side effects.

Disclosure of Invention

The present invention provides a compound of formula (I), an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, useful as a selective inhibitor of BCL-2,

wherein:

X ¹ a 6 membered saturated heterocyclic group selected from optionally substituted containing one or two heteroatoms selected from N, O, S, wherein the optional substituent is selected from 4 membered saturated heterocyclic groups containing one or two heteroatoms selected from N, O, S; preferably X ¹ A 6 membered saturated heterocyclic group selected from optionally substituted containing one or two heteroatoms selected from N, O, wherein the optional substituent is selected from oxetanyl; further preferably, X ¹ Selected from 1, 4-dioxanyl, tetrahydropyranyl, N-oxetanyl piperidinyl, N-oxetanyl morpholinyl; most preferably, X ¹ Selected from (S) -1, 4-dioxane-2-yl, (R) -1, 4-dioxane-2-yl, tetrahydropyran-4-yl, 1- (oxetan-3-yl) piperidin-4-yl, (S) -4- (oxetan-3-yl) morpholin-2-yl;

X ² a 5-6 membered heterocyclic group selected from the group consisting of containing one or two N atoms; wherein the ring is optionally substituted with one or two C1-C4 alkyl groups; preferably X ² A 6 membered heterocyclic group selected from one or two N atoms; wherein the ring is optionally substituted with one or two C1-C4 alkyl groups; further preferably, X ² Selected from the group consisting of

Wherein the ring is optionally substituted with a C1-C4 alkyl group; most preferably, X ² Selected from the group consisting of

R ⁰ Selected from hydrogen, halogen; preferably, R ⁰ Selected from hydrogen, fluorine, chlorine; most preferably, R ⁰ Selected from hydrogen, fluorine;

R ¹ 、R ² 、R ³ 、R ⁴ each independently selected from hydrogen, C1-C6 alkyl, wherein R ¹ And R is ² Or R is ³ And R is ⁴ Can form 3-6 membered cycloalkyl together with the attached carbon atom; preferably, R ¹ 、R ² 、R ³ 、R ⁴ Each independently selected from hydrogen, C1-C4 alkyl, wherein R ¹ And R is ² Or R is ³ And R is ⁴ Can form 3-4 membered cycloalkyl together with the attached carbon atom; further preferably, R ¹ 、R ² 、R ³ 、R ⁴ Each independently selected from hydrogen or methyl and R ¹ 、R ² 、R ³ 、R ⁴ Not all hydrogen, wherein R ¹ And R is ² Or R is ³ And R is ⁴ Can form a cyclopropyl group together with the attached carbon atom; most preferably, R ¹ 、R ² 、R ³ 、R ⁴ Each independently selected from hydrogen or methyl and R ¹ 、R ² 、R ³ 、R ⁴ Not all hydrogen, wherein R ³ And R is ⁴ Cyclopropyl may be formed together with the attached carbon atom.

Preferably, the present invention relates to a compound of formula (I) as described above, an isomer, a prodrug, a solvate, a stable isotope derivative or a pharmaceutically acceptable salt thereof, wherein:

X ¹ a 6 membered saturated heterocyclic group selected from optionally substituted containing one or two heteroatoms selected from N, O, wherein the optional substituent is selected from oxetanyl;

X ² A 6 membered heterocyclic group selected from one or two N atoms; wherein the ring is optionally substituted with one or two C1-C4 alkyl groups;

R ⁰ selected from hydrogen, halogen;

R ¹ 、R ² 、R ³ 、R ⁴ each independently selected from hydrogen, C1-C4 alkyl, wherein R ¹ And R is ² Or R is ³ And R is ⁴ Can together with the attached carbon atom form a 3-to 4-membered cycloalkyl group.

Further preferred, the present invention relates to a compound of formula (I) as described hereinbefore, isomers, prodrugs, solvates, stable isotope derivatives or pharmaceutically acceptable salts thereof, wherein:

X ² is that

Wherein the ring is optionally substituted with a C1-C4 alkyl group;

R ⁰ selected from hydrogen, fluorine, chlorine;

X ¹ selected from 1, 4-dioxanyl, tetrahydropyranyl, N-oxetanyl piperidinyl, N-oxetanyl morpholinyl;

X ² Is that

Wherein the ring is optionally substituted with a C1-C4 alkyl group;

R ⁰ selected from hydrogen, fluorine, chlorine;

R ¹ 、R ² 、R ³ 、R ⁴ each independently selected from hydrogen or methyl and R ¹ 、R ² 、R ³ 、R ⁴ Not all hydrogen, wherein R ¹ And R is ² Or R is ³ And R is ⁴ Cyclopropyl may be formed together with the attached carbon atom.

Still further preferred, the present invention relates to a compound of formula (I) as hereinbefore described, isomers, prodrugs, solvates, stable isotope derivatives or pharmaceutically acceptable salts thereof, wherein:

X ² selected from the group consisting of

R ⁰ Selected from hydrogen, fluorine;

X ¹ selected from (S) -1, 4-dioxane-2-yl, (R) -1, 4-dioxane-2-yl, tetrahydropyran-4-yl, 1- (oxetan-3-yl) piperidin-4-yl, (S) -4- (oxetan-3-yl) morpholin-2-yl;

X ² selected from the group consisting of

R ⁰ Selected from hydrogen, fluorine;

R ¹ 、R ² 、R ³ 、R ⁴ Each independently selected from hydrogen or methyl and R ¹ 、R ² 、R ³ 、R ⁴ Not all hydrogen, wherein R ³ And R is ⁴ Cyclopropyl may be formed together with the attached carbon atom.

Most preferably, the present invention relates to a compound of formula (I) as described previously, an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, selected from:

the invention also relates to the use of a compound of formula (I), an isomer, a prodrug, a solvate, a stable isotope derivative or a pharmaceutically acceptable salt thereof according to any of the embodiments of the invention for the manufacture of a medicament for use as a BCL-2 inhibitor.

The invention also relates to the use of a compound of formula (I) or an isomer, prodrug, solvate, stable isotope derivative or a pharmaceutically acceptable salt thereof according to any of the embodiments of the invention for the manufacture of a medicament for the treatment or prophylaxis of BCL-2 mediated related diseases such as tumors selected from hematological malignancies, lung cancer, breast cancer, ovarian cancer, prostate cancer, rectal cancer, pancreatic cancer, glioma, especially acute lymphoblastic leukemia.

The invention further relates to a pharmaceutical composition comprising a compound of formula (I) or an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof as described in any of the embodiments of the invention, optionally one or more other BCL-2 inhibitors, and one or more pharmaceutically acceptable carriers, diluents and excipients.

The invention also relates to the use of a pharmaceutical composition according to the invention for the preparation of a medicament for the treatment or prevention of a related disease mediated by BCL-2, such as a tumor selected from hematological malignancies, lung cancer, breast cancer, ovarian cancer, prostate cancer, rectal cancer, pancreatic cancer, brain glioma, in particular acute lymphoblastic leukemia.

The present invention also relates to a method of treating or preventing a BCL-2 mediated related disease comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any one of the embodiments of the present invention or an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof; or a pharmaceutical composition according to the invention, for example a tumor selected from hematological malignancies, in particular acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, prostate cancer, rectal cancer, pancreatic cancer, glioma.

A further aspect of the invention relates to a compound as described in any one of the embodiments of the invention, or an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, for use in the treatment or prophylaxis of a disease associated with BCL-2, such as a tumour selected from hematological malignancies, lung cancer, breast cancer, ovarian cancer, prostate cancer, rectal cancer, pancreatic cancer, glioma, in particular acute lymphoblastic leukemia.

Another aspect of the invention relates to a pharmaceutical composition comprising a compound of formula (I) or an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof as described in any of the embodiments of the invention, optionally one or more other BCL-2 inhibitors, and one or more pharmaceutically acceptable carriers, diluents and excipients for the treatment or prophylaxis of a related disease mediated by BCL-2, such as a tumor selected from the group consisting of hematological malignancies, lung cancer, breast cancer, ovarian cancer, prostate cancer, rectal cancer, pancreatic cancer, brain glioma, especially acute lymphoblastic leukemia.

Another aspect of the present invention relates to a compound of formula (I) or an isomer, prodrug, solvate, stable isotope derivative or a pharmaceutically acceptable salt thereof as described in any one of the embodiments of the present invention for use in the treatment and/or prevention of BCL-2 mediated related diseases. Such BCL-2 mediated related diseases as e.g. tumors selected from hematological malignancies, lung cancer, breast cancer, ovarian cancer, prostate cancer, rectal cancer, pancreatic cancer, brain glioma, especially acute lymphoblastic leukemia.

According to the present invention, the drug may be any pharmaceutical dosage form including, but not limited to, tablets, capsules, solutions, lyophilized formulations, injections.

The pharmaceutical formulations of the present invention may be administered in dosage unit form containing a predetermined amount of active ingredient per dosage unit. Such units may contain, for example, from 0.5 mg to 1 g, preferably from 1 mg to 700 mg, particularly preferably from 5 mg to 300 mg of a compound of the invention, or the pharmaceutical formulation may be administered in dosage unit form containing a predetermined amount of active ingredient per dosage unit, depending on the disorder being treated, the method of administration and the age, weight and condition of the patient. Preferred dosage unit formulations are those containing a daily dose or divided dose, or a corresponding fraction thereof, of the active ingredient as indicated above. In addition, this type of pharmaceutical formulation may be prepared using methods well known in the pharmaceutical arts.

The pharmaceutical formulations of the invention may be adapted for administration by any desired suitable method, for example by oral (including buccal or sublingual), rectal, nasal, topical (including buccal, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) methods. Such formulations may be prepared using all methods known in the pharmaceutical arts by, for example, combining the active ingredient with one or more excipients or one or more adjuvants.

Preparation flow

The invention also provides a method for preparing the compound.

Scheme 1

R ⁰ And X is ¹ Is as defined above;

the first step:

dissolving the compound (I) in chlorosulfonic acid, reacting for 10-20 hours under the condition of heating in an oil bath (120-150 ℃), cooling to room temperature, quenching with ice water, extracting with ethyl acetate, dissolving a crude product obtained by organic phase drying and concentrating in anhydrous tetrahydrofuran, dropwise adding ammonia water at a low temperature (-80-60 ℃), continuously stirring for 1-5 hours, and acidifying with hydrochloric acid to obtain the compound (II);

and a second step of:

dissolving the compound (II) and corresponding amine in a solvent (such as acetonitrile), adding a base (such as triethylamine or diisopropylethylamine, etc.), and stirring at 25-60 ℃ for 10-20 hours under the protection of inert gas (such as nitrogen or argon) to obtain a compound (III);

scheme 2

R ¹ 、R ² 、R ³ 、R ⁴ Is as defined above;

the first step:

dropwise adding phosphorus oxychloride into a dichloromethane solution of N, N-dimethylformamide in an ice bath under the protection of nitrogen, stirring at room temperature for 30 minutes after dropwise adding, cooling to 0 ℃ again, dropwise adding a dichloromethane solution of a compound (IV), and reacting at room temperature-60 ℃ for 10-24 hours to obtain a compound (V);

and a second step of:

adding a compound (V), p-chlorobenzeneboronic acid, alkali such as potassium carbonate, a phase transfer catalyst such as tetra-n-butyl ammonium bromide and a catalyst such as palladium acetate into a solvent such as water under the protection of nitrogen, vacuumizing the system, replacing the system with nitrogen for three times, and heating to 40-100 ℃ for reacting for 2-10 hours to obtain a compound (VI);

And a third step of:

dissolving the compound (VI) in a solvent such as tetrahydrofuran and methanol, adding a reducing agent such as sodium borohydride, and stirring at room temperature for 1-5 hours to obtain a compound (VII);

fourth step:

dissolving the compound (VII) in a solvent such as dichloromethane, adding a chloro reagent such as thionyl chloride, and stirring at room temperature for 10-24 hours to obtain the compound (VIII);

scheme 3

R ⁰ 、R ¹ 、R ² 、R ³ 、R ⁴ And X is ¹ Is as defined above;

the first step:

dissolving compound (IX) (synthetic reference: WO 2017212431) (0.51 g,2.00 mmol) and tert-butyl (R) -3-methylpiperazine-1-carboxylate in a solvent (e.g., dimethyl sulfoxide), adding a base such as dipotassium hydrogen phosphate at room temperature, and stirring at 90-120deg.C for 12-48 hours to obtain compound (X);

and a second step of:

adding the compound (X) into a 1, 4-dioxane solution of acid such as hydrogen chloride, and stirring at room temperature for 1-3 hours to obtain a compound (XI);

and a third step of:

dissolving the compound (XI) and the compound (VIII) in a solvent such as acetonitrile, adding a base such as N, N-diisopropylethylamine, heating to 60-90 ℃ and stirring for 2-8 hours to obtain a compound (XII);

fourth step:

dissolving compound (XII) in solvent such as 2-butanol, adding alkali such as sodium hydroxide, heating to 90-120deg.C, stirring for 12-36 hr, and acidifying with acid such as hydrochloric acid to obtain compound (XIII);

Fifth step:

dissolving a compound (XIII) and a compound (III) in a solvent such as dichloromethane, and stirring at room temperature for 12-36 hours to obtain a compound (XIV) by using a condensing agent such as 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and a base such as 4-dimethylaminopyridine;

scheme 4

R ⁰ 、R ¹ 、R ² 、R ³ 、R ⁴ And X is ¹ Is as defined above;

the first step:

dissolving a compound (XV) (synthetic reference: journal of Organic Chemistry,84 (8), 4814-4829; 2019) (0.51 g,2.00 mmol) and (R) -2-methylpiperazine in a solvent (such as dimethyl sulfoxide), adding a base such as N, N-diisopropylethylamine at room temperature, and stirring at 50-100deg.C for 12-24 hours to give a compound (XVI);

and a second step of:

dissolving the compound (XVI) and the compound (VIII) in a solvent such as acetonitrile, adding a base such as N, N-diisopropylethylamine, heating to 50-90 ℃ and stirring for 8-24 hours to obtain the compound (XVII);

and a third step of:

dissolving the compound (XVII) in a solvent such as water, methanol and tetrahydrofuran, adding a base such as lithium hydroxide, heating to 50-90 ℃ and stirring for 1-5 hours, and acidifying with an acid such as hydrochloric acid to obtain the compound (XVIII);

fourth step:

compound (XVIII), compound (III), condensing agent such as 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride, base such as 4-dimethylaminopyridine are dissolved in solvent such as dichloromethane and stirred at room temperature for 12 to 36 hours to give compound (XIX).

Detailed Description

Definition of the definition

Unless stated to the contrary, the following terms used in the specification and claims have the following meanings. The groups not specifically defined in the present invention have meanings commonly represented in the art as known to those skilled in the art.

The expression "Cx-Cy" as used in the present invention means a range of carbon atom numbers, wherein x and y are both integers, for example, C3-C8 cycloalkyl represents cycloalkyl having 3 to 8 carbon atoms, -C0-C2 alkyl represents alkyl having 0 to 2 carbon atoms, wherein-C0 alkyl means a chemical single bond.

In the present invention, the term "alkyl" refers to saturated aliphatic hydrocarbon groups, including straight and branched chain groups of 1 to 20 carbon atoms, which may be, for example, straight and branched chain groups of 1 to 18 carbon atoms, 1 to 12 carbon atoms, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1-dimethylpropyl, 1, 2-dimethylpropyl, 2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1, 2-trimethylpropyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, and various branched isomers thereof, and the like. The alkyl group may be optionally substituted or unsubstituted.

In the present invention, the term "cycloalkyl" refers to a saturated monocyclic or polycyclic cyclic hydrocarbon group comprising 3 to 12 ring atoms, which may be, for example, 3 to 12, 3 to 10, 3 to 8 or 3 to 6 ring atoms, or may be a 3, 4, 5, 6 membered ring. Non-limiting examples of monocyclic ring groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, and the like. Cycloalkyl groups may be optionally substituted or unsubstituted.

In the present invention, the term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon group comprising 3 to 20 ring atoms, which may be, for example, 3 to 16, 3 to 12, 3 to 10, 3 to 8 or 3 to 6 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen or heteroatoms of S (O) m (where m is an integer from 0 to 2), but excluding the ring portion of-O-, -O-S-or-S-, the remaining ring atoms being carbon. Preferably 3 to 12 ring atoms, of which 1 to 4 are heteroatoms, more preferably the heterocyclyl ring contains 3 to 10 ring atoms, more preferably 3 to 8 ring atoms, most preferably 5-membered ring or 6-membered ring, of which 1 to 4 are heteroatoms, more preferably 1 to 3 are heteroatoms, most preferably 1 to 2 are heteroatoms. Non-limiting examples of monocyclic heterocyclyl groups include oxetanyl, pyrrolidinyl, piperidinyl, 4-piperidinyl, piperazinyl, 1, 4-dioxanyl, morpholinyl, 2-morpholinyl, 4-morpholinyl, thiomorpholinyl, pyranyl, tetrahydropyranyl, 4-tetrahydropyranyl, homopiperazinyl, dioxanyl, 2-dioxanyl, and the like. Polycyclic heterocyclyl groups include spiro, fused and bridged heterocyclic groups. The heterocyclyl group may be optionally substituted or unsubstituted.

In the present invention, the term "heterocyclylene" refers to a substituted or unsubstituted heterocyclic group having two terminal monovalent radical cores, resulting from the removal of one hydrogen atom from each of the two terminal atoms; the heterocyclic group has the meaning as described above. Non-limiting examples of "heterocyclylene" include pyrrolidinylene, piperidinyl, piperazinylene, morpholinylene, and the like.

In the present invention, the term "halogen" refers to fluorine, chlorine, bromine or iodine.

In the present invention, "optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs or does not. For example, "a heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may be, but is not necessarily, present, and the description includes cases where the heterocyclic group is substituted with an alkyl group and cases where the heterocyclic group is not substituted with an alkyl group.

In the present invention, "substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms in the group are independently substituted with a corresponding number of substituents. It goes without saying that substituents are only in their possible chemical positions, and that the person skilled in the art is able to determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups having free hydrogen may be unstable when bound to carbon atoms having unsaturated (e.g., olefinic) bonds.

Such substituents include, but are not limited to, the various groups described previously.

The claimed compounds include not only the compounds themselves, but also isomers, prodrugs, solvates, stable isotopic derivatives of the compounds or pharmaceutically acceptable salts thereof.

The term "pharmaceutical composition" as used herein means a mixture containing one or more of the isomers, prodrugs, solvates, stable isotope derivatives, or pharmaceutically acceptable salts thereof, and other chemical components of the compounds of the present invention. Other components such as pharmaceutically acceptable carriers, diluents and excipients. The purpose of the pharmaceutical composition is to promote the administration to organisms, facilitate the absorption of active ingredients and thus exert biological activity.

The term "comprising" when used in the specification is comprised of ….

The term "room temperature" as used herein means 15-30deg.C.

The "stable isotope derivatives" of the present invention include: an isotopically substituted derivative in which any hydrogen atom in the formula (I) is substituted with 1 to 5 deuterium atoms, an isotopically substituted derivative in which any carbon atom in the formula (I) is substituted with 1 to 3 carbon 14 atoms, or an isotopically substituted derivative in which any oxygen atom in the formula (I) is substituted with 1 to 3 oxygen 18 atoms.

The "pharmaceutically acceptable salts" of the present invention are discussed in Berge, et al, "Pharmaceutically acceptable salts," j.pharm.sci.,66,1-19 (1977) and are readily apparent to pharmaceutical chemists that are substantially non-toxic and provide desirable pharmacokinetic properties, palatability, absorption, distribution, metabolism, excretion, and the like.

Pharmaceutically acceptable salts of the invention can be synthesized by general chemical methods.

In general, salts can be prepared by reacting the free base or acid with an equivalent stoichiometric or excess of an acid (inorganic or organic) or base in a suitable solvent or solvent composition.

The term "prodrug" as used herein refers to a compound that is converted to the original active compound after metabolism in vivo. Typically, prodrugs are inactive substances or less active than the active parent compound, but may provide ease of handling, administration or improved metabolic characteristics.

The term "isomer" as used herein refers to the tautomers, meso, racemates, enantiomers, diastereomers, mixtures thereof, and the like, of the compounds of formula (I) according to the invention. All such isomers, including stereoisomers, geometric isomers are encompassed by the present invention. The geometric isomers include cis and trans isomers.

The term "solvate" as used herein refers to an association of one or more solvent molecules with a compound of the invention or a salt thereof. Examples of solvents that form pharmaceutically acceptable solvates include, but are not limited to, water, isopropanol, ethanol, methanol, ethyl acetate, acetic acid, and the like.

The invention includes any polymorph of the compound or salt thereof, and any hydrate or other solvate.

In the present invention, the term "patient" generally refers to a mammal, especially a human.

In the present invention, the term "tumor" includes benign tumors and malignant tumors, such as cancers.

In the present invention, the term "cancer" includes various tumors mediated by BCL-2, including but not limited to hematological malignancies, lung cancer, breast cancer, ovarian cancer, prostate cancer, rectal cancer, pancreatic cancer, brain glioma, among others, acute lymphoblastic leukemia.

In the present invention, the term "therapeutically effective amount" is meant to include an amount of a compound of the present invention that is effective to treat or prevent a related disorder mediated by BCL-2.

Examples

The invention is further illustrated by means of the following examples, which are not intended to limit the scope of the invention. The experimental methods, in which specific conditions are not noted in the following examples, were selected according to conventional methods and conditions, or according to the commercial specifications.

The structures of all compounds of the invention can be identified by nuclear magnetic resonance (1H NMR) and/or mass spectrometry detection (MS).

¹ H NMR chemical shifts (δ) are reported in PPM (parts per million ). NMR was performed by Bruker AVANCE III-400MHz spectrometer. Suitable solvents are selected from deuterated chloroform (CDCl) ₃ ) Deuterated methanol (CD) ₃ OD), deuterated dimethyl sulfoxide (DMSO-d) ⁶ ) And the like, tetramethylsilane as an internal standard (TMS).

Low resolution Mass Spectrometry (MS) was determined by an Agilent 1260 HPLC/6120 mass spectrometer using Agilent ZORBAX XDB-C18, 4.6X105 mm,3.5 μm.

Gradient elution condition one: 0:95% solvent A1 and 5% solvent B1, 1-2:5% solvent A1 and 95% solvent B1; 2.01-2.50:95% solvent A1 and 5% solvent B1. The percentage is the volume percentage of a certain solvent to the total solvent volume. Solvent A1:0.01% formic acid aqueous solution; solvent B1:0.01% formic acid in acetonitrile; the percentage is the volume percentage of solute in the solution.

The thin-layer silica gel plate is a tobacco stand yellow sea HSGF254 or Qingdao GF254 silica gel plate. Column chromatography generally uses 100-200 or 200-300 mesh silica gel of yellow sea as carrier.

Preparative liquid chromatography (prep-HPLC) using Waters SQD2 mass spectrometry directed to a high pressure liquid chromatography separator, XBridge-C18;30X 150mm preparation column, 5 μm;

The method comprises the following steps: acetonitrile-water (0.2% formic acid), flow rate 25 mL/min; the second method is as follows: acetonitrile-water (0.8% ammonium bicarbonate), flow rate 25 mL/min;

the known starting materials of the present invention may be synthesized using or following methods known in the art, or may be purchased from Acros Organics, aldrich Chemical Company, shaoshan chemical technology (Accela ChemBio Inc), shanghai Piobtained medicine, shanghai Ala Ding Huaxue, shanghai Michelson chemistry, carbofuran chemistry, anaesthetic and chemistry, among others. Venetoclax was purchased from chemical technology Co., ltd.

In the examples, if no special description exists, the solvent used in the reaction is anhydrous solvent, wherein the anhydrous tetrahydrofuran is commercial tetrahydrofuran, sodium block is used as a water scavenger, benzophenone is used as an indicator, reflux is carried out to the solution under the protection of argon gas to form bluish violet, distillation and collection are carried out, the solution is stored at room temperature under the protection of argon gas, other anhydrous solvents are purchased from Anaesthetic and chemical and carbofuran chemistry, and all transfer and use of the anhydrous solvents are carried out under the protection of argon gas if no special description exists.

In the examples, unless otherwise specified, the reaction was carried out under an argon atmosphere or a nitrogen atmosphere.

An argon or nitrogen atmosphere means that the reactor flask is connected to a balloon of argon or nitrogen of about 1L volume.

The hydrogen atmosphere is defined as the reaction flask being connected to a balloon of hydrogen gas of about 1L volume.

The hydrogenation reaction is usually vacuumized, filled with hydrogen and repeatedly operated for 3 times.

In the examples, the reaction temperature was room temperature and the temperature range was 15℃to 30℃unless otherwise specified.

The progress of the reaction in the examples was monitored by Thin Layer Chromatography (TLC) using a system of developing agents of a: methylene chloride and methanol systems; b: petroleum ether and ethyl acetate systems. The volume ratio of the solvent is adjusted according to the polarity of the compound.

The system of eluent for column chromatography and the system of developing agent for thin layer chromatography used for purifying the compound include a: methylene chloride and methanol systems; b: petroleum ether and ethyl acetate systems. The volume ratio of the solvent is adjusted according to the polarity of the compound, and can be adjusted by adding a small amount of triethylamine, an acidic or alkaline reagent and the like.

Intermediate 1

3-fluoro-5-nitro-4- (((tetrahydro-2H-pyran-4-yl) methyl) amino) benzenesulfonamide

First step

3, 4-difluoro-5-nitrobenzenesulfonamide

1, 2-difluoro-3-nitrobenzene (10.00 g,62.89 mmol) was dissolved in chlorosulfonic acid (21 mL), heated to 150℃and stirred at reflux for 10 hours. Cooled to room temperature, and a saturated aqueous sodium hydrogencarbonate solution was added to the reaction mixture in an ice bath to adjust the pH to about 7. The mixture was extracted with methylene chloride (100 mL. Times.3), and the organic phase was washed with saturated brine (100 mL. Times.2), dried over anhydrous sodium sulfate, and concentrated by filtration to give the crude 3, 4-difluoro-5-nitrobenzenesulfonyl chloride. Isopropanol (200 mL) and aqueous ammonia (5 mL, 37%) were added to a 1000mL three-necked flask and stirred at-78deg.C for 10 minutes, the resulting crude 3, 4-difluoro-5-nitrobenzenesulfonyl chloride was dissolved in isopropanol (30 mL), slowly dropped into the above mixture of isopropanol and aqueous ammonia at-78deg.C, stirred at-78deg.C for two hours after the completion of the dropping, and diluted hydrochloric acid (1N) was added to adjust the pH of the system to about 6. And (3) heating to room temperature, concentrating under reduced pressure to remove most of isopropanol solvent, adding pure water into the solvent, separating out solid, and filtering to obtain a solid crude product. The crude product was purified by beating with methylene chloride to give the target product 3, 4-difluoro-5-nitrobenzenesulfonamide (4.90 g, yellow solid). Yield: 33%.

¹ H NMR(400MHz，DMSO-d ₆ )δ8.38-8.36(m，1H)，8.29-8.26(m，1H)，7.84(s，2H)。

Second step

The compound 3, 4-difluoro-5-nitrobenzenesulfonamide (1.55 g,6.51 mmol), (tetrahydro-2H-pyran-4-yl) methylamine (0.89 g,7.73 mmol), N-diisopropylethylamine (3.91 g,30.31 mmol) and acetonitrile (20.0 mL) were mixed. Stirred at 40℃for 2 hours. The solvent was dried, the mixture was quenched with 50mL of water and extracted with ethyl acetate (60 mL. Times.2). The combined organic phases were washed with saturated brine (50 mL. Times.3). The organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the crude product was obtained by desolventizing under reduced pressure. Column chromatography purification (petroleum ether/ethyl acetate=1:1) afforded the target product 3-fluoro-5-nitro-4- (((tetrahydro-2H-pyran-4-yl) methyl) amino) benzenesulfonamide (1.66 g, yellow solid). Yield: 76%.

MS m/z(ESI)：334[M+1]；

¹ H NMR(400MHz，DMSO-d ₆ )δ8.33-8.30(m，2H)，7.76-7.74(m，1H)，7.45(s，2H)，3.86-3.84(m，2H)，3.50-3.45(m，2H)，3.29-3.23(m，3H)，1.59-1.57(m，2H)，1.25-1.20(m，2H)。

The synthesis step of intermediate 2 refers to intermediate 1.

Intermediate 3

3-fluoro-5-nitro-4- (((1- (3-oxetanyl) piperidin-4-yl) methyl) amino) benzenesulfonamide

First step

3-fluoro-5-nitro-4- ((piperidin-4-ylmethyl) amino) benzenesulfonamide

The compound 4- (((2-fluoro-6-nitro-4-sulfamoylphenyl) amino) methyl) piperidine-1-carboxylic acid tert-butyl ester (synthesis procedure for synthesis of reference intermediate 1) (0.49 g,1.12 mmol) was mixed with trifluoroacetic acid (3 mL), dichloromethane (9 mL) and stirred at room temperature for 0.5 hours. The solvent was dried, the pH of the mixture was adjusted to neutrality with triethylamine, and the solvent was again dried to give crude 3-fluoro-5-nitro-4- ((piperidin-4-ylmethyl) amino) benzenesulfonamide (0.35 g, yellow oil). This mixture was used in the next reaction without purification. MS m/z (ESI): 333 m+1.

Second step

The compound 3-fluoro-5-nitro-4- ((piperidin-4-ylmethyl) amino) benzenesulfonamide (0.35 g,1.05 mmol) and oxetan-3-one (0.23 g,3.15 mmol), sodium cyanoborohydride (0.23 g,5.23 mmol) and methanol (8 mL) were mixed and stirred at ambient temperature for 1.5 hours. The mixture was quenched with 10mL of water and extracted with ethyl acetate (10 mL. Times.2). The combined organic phases were washed with saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, dried by spin-drying, and the residue was purified by column chromatography (petroleum ether/ethyl acetate=3:1) to give the desired product 3-fluoro-5-nitro-4- (((1- (3-oxetanyl) piperidin-4-yl) methyl) amino) benzenesulfonamide (0.32 g, yellow solid). Yield: 78%.

MS m/z(ESI)：389[M+1]；

¹ H NMR(400MHz，CD ₃ OD)δ8.47-8.46(m，1H)，7.72-7.70(m，1H)，4.68-4.65(m，2H)，4.60-4.59(m，2H)，3.57-3.55(m，2H)，3.49-3.45(m，1H)，2.83-2.80(m，2H)，1.89-1.78(m，3H)，1.42-1.20(m，4H)。

Intermediate 4

(R) -4 '-chloro-6- (chloromethyl) -3-methyl-2, 3,4, 5-tetrahydro-1, 1' -biphenyl

First step

(R) -2-chloro-4-methylcyclohex-1-ene-1-carbaldehyde

N, N-dimethylformamide (0.59 g,8.00 mmol) was dissolved in methylene chloride (10 mL), the temperature was lowered to 0℃in an ice bath, phosphine oxide trichloride (0.92 g,6.00 mmol) was added dropwise, and after stirring for 30 minutes with heat preservation, the mixture was naturally warmed to room temperature and stirring was continued for 3 hours. Cooled to 0 ℃, a solution of (R) -3-methylcyclohexyl-1-one (synthetic reference: tetrahedron 73 (2017) 3202-3212) (0.45 g,4.0 mmol) in dichloromethane (5 mL) was slowly added, and the reaction was slowly warmed to room temperature and stirred for 18 hours. The reaction mixture was quenched with saturated sodium bicarbonate solution (10 mL), concentrated to remove dichloromethane, ethyl acetate (25 mL) and water (15 mL) were added, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (25 mL. Times.2), the combined organic phases were washed with saturated brine (20 mL. Times.2), dried over anhydrous sodium sulfate, and concentrated by filtration to give the title compound (R) -2-chloro-4-methylcyclohex-1-ene-1-carbaldehyde (0.45 g, pale yellow liquid), crude product.

MS m/z(ESI)：159&161[M+1]；

¹ H NMR(400MHz，CDCl ₃ )δ10.20(s，0.8H)，10.18(s，0.2H)，2.63-2.47(m，2H)，2.29-2.26(m，1H)，2.24-2.15(m，1H)，1.90-1.77(m，2H)，1.27-1.21(m，1H)，1.03(d，J＝6.4Hz，3H)。

Second step

(R) -4 '-chloro-5-methyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-carbaldehyde

A single-necked flask was charged with (R) -2-chloro-4-methylcyclohex-1-ene-1-carbaldehyde (0.45 g,2.80 mmol), p-chlorobenzeneboronic acid (0.44 g,2.80 mmol), tetrabutylammonium bromide (0.91 g,2.80 mmol), potassium carbonate (1.17 g,8.50 mmol), palladium acetate (0.16 g,0.70 mmol) and water (15 mL) in this order, the reaction system was replaced with nitrogen gas for 3 times, and heated to 45℃for 4 hours. The reaction solution was cooled to room temperature, ethyl acetate (25 mL) was added, the organic phase was separated, the aqueous phase was extracted with ethyl acetate (25 mL), the organic phases were combined, washed with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and the residue was purified by column chromatography (petroleum ether: ethyl acetate=96:4) to give the objective compound (R) -4 '-chloro-5-methyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-carbaldehyde (0.45 g, pale yellow oily substance) in 67.5% yield.

MS m/z(ESI)：235&237[M+1]；

¹ H NMR(400MHz，CDCl ₃ )δ9.41(s，0.9H)，9.37(s，0.1H)，7.30-7.24(m，2H)，7.10-7.08(m，2H)，2.54-2.47(m，2H)，2.16-2.03(m，2H)，1.82-1.75(m，2H)，1.21-1.15(m，1H)，0.96(d，J＝6.4Hz，3H)。

Third step

(R) - (4 '-chloro-5-methyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methanol

(R) -4 '-chloro-5-methyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-carbaldehyde (0.23 g,1.00 mmol) was dissolved in tetrahydrofuran (2 mL), and sodium borohydride (57 mg,1.50 mmol) was added at 0deg.C and stirred at room temperature for 2 hours. The reaction was quenched by dropwise addition of saturated aqueous ammonium chloride (10 mL), extracted with ethyl acetate (10 mL), the organic phase was washed with saturated brine (10 mL), dried, filtered, and the filtrate concentrated to give crude (R) - (4 '-chloro-5-methyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methanol (0.24 g, pale yellow oil) which was used directly in the next step without purification. MS m/z (ESI): 237&239[ M-17].

Fourth step

(R) - (4 '-chloro-5-methyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methanol (0.24 g,1.00 mmol) was dissolved in dichloromethane (2 mL), and thionyl chloride (0.24 g,1.50 mmol) was added at room temperature and stirred for 1 hour. The reaction was quenched by dropwise addition of saturated aqueous sodium bicarbonate (10 mL), extracted with dichloromethane (10 mL), the organic phase was washed with saturated brine (10 mL), dried, filtered, the filtrate concentrated, and the reaction solution was spun-dried to give the crude (R) -4 '-chloro-6- (chloromethyl) -3-methyl-2, 3,4, 5-tetrahydro-1, 1' -biphenyl (0.26 g, pale yellow oil) which was used directly in the next step without purification.

Synthesis of intermediate 5-7 reference the synthesis of intermediate 4.

Intermediate 8

5- (chloromethyl) -6- (4-chlorophenyl) -8, 8-dimethyl spiro [2.5] oct-5-ene

First step

7, 7-dimethyl-8-methylene-1, 4-dioxaspiro [4.5] decane

The compound methyltriphenylphosphine bromide (14.00 g,39.20 mmol) was added to anhydrous tetrahydrofuran (120 mL), cooled to-40℃and then added dropwise to a solution of n-butyllithium in n-hexane (2.5M, 39.2mmol,15.6 mL) under nitrogen, after the addition, the mixture was allowed to warm to room temperature and stirred for 1 hour. 7, 7-dimethyl-1, 4-dioxaspiro [4.5] decan-8-one (6.00 g,32.60 mmol) was then added dropwise and reacted overnight at room temperature. The reaction mixture was quenched with saturated aqueous ammonium chloride (200 mL), the aqueous phase was extracted with ethyl acetate (100 mL. Times.2), the organic phases were combined, washed with saturated brine (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure. The residue was chromatographed on silica gel (0-10% ethyl acetate/petroleum ether) to give the product 7, 7-dimethyl-8-methylene-1, 4-dioxaspiro [4.5] decane (3.80 g, colorless oil). Yield: 64%.

¹ H NMR(400MHz，CDCl ₃ )δ4.73(s，2H)，3.98-3.94(m，4H)， 2.43-2.36(m，2H)，1.73-1.68(m，2H)，1.62(s，2H)，1.16(s，6H)。

Second step

4, 4-dimethyl-7, 10-dioxadispiro [2.2.4 ⁶ .2 ³ ]Dodecane (dodecane)

The compound 7, 7-dimethyl-8-methylene-1, 4-dioxaspiro [4.5 ]]Decane (0.90 g,4.94 mmol) was dissolved in methylene chloride (20 mL), and a toluene solution of diethyl zinc (2M, 14.80mmol,7.4 mL) was added dropwise under nitrogen at 0℃and stirred for 0.5 hours after the addition. Chloroiodomethane (5.20 g,29.60 mmol) was then added dropwise and stirred overnight at room temperature. The reaction was quenched with saturated aqueous ammonium chloride (30 mL), the organic phase was separated, the aqueous phase extracted with dichloromethane (30 mL. Times.2), and the combined organic phases were washed with water (30 mL. Times.2) and saturated brine (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure to give the crude product 4, 4-dimethyl-7, 10-dioxadispiro [2.2.4 ] ⁶ .2 ³ ]Dodecane (1.50 g, yellow oil). Crude product.

¹ H NMR(400MHz，CDCl ₃ )δ3.96-3.91(m，4H)，1.69-1.62(m，2H)，1.55(s，2H)，1.44-1.40(m，2H)，0.83(s，6H)，0.50-0.46(m，2H)，0.11-0.08(m，2H)。

Third step

4, 4-Dimethylspiro [2.5] octan-6-one

The compound 4, 4-dimethyl-7, 10-dioxadispiro [2.2.4 ⁶ .2 ³ ]Dodecane (1.50 g, crude product, 4.94 mmol) was dissolved in tetrahydrofuran hydrochloride mixture (2M, 14.00mmol,7 mL) and stirred at room temperature for 2 hours. The reaction mixture was quenched by addition of saturated aqueous sodium bicarbonate (30 mL). The mixture was extracted with ethyl acetate (30 mL), and the organic phase was washed with water (30 mL. Times.2) and saturated brine (30 mL), respectively. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 4, 4-dimethyl spiro [2.5] as a crude product ]Octan-6-one (0.70 g, yellow oil). The strictly rate: 93.2%. MS m/z (ESI): 153[ M+1]]。

Fourth step

6-chloro-8, 8-dimethyl-spiro [2.5] oct-5-ene-5-carbaldehyde

Phosphorus oxychloride (1.10 g,6.90 mmol) was added dropwise to a solution of ice-bath N, N-dimethylformamide (0.67 g,9.20 mmol) in methylene chloride (30 mL) under nitrogen protection, after the addition was completed, the mixture was stirred at room temperature for 30 minutes, cooled again to 0℃and a solution of 4, 4-dimethyl spiro [2.5] octan-6-one (0.70 g,4.60 mmol) in methylene chloride (5 mL) was added dropwise thereto, and the mixture was reacted overnight at room temperature. The reaction was quenched with water (30 mL), and the organic phase was separated and washed with saturated sodium bicarbonate solution (30 mL) and saturated brine (30 mL), respectively. The organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (100% petroleum ether) to give 6-chloro-8, 8-dimethyl spiro [2.5] oct-5-ene-5-carbaldehyde (0.34 g, pale yellow oily substance). Yield: 37.2%. MS m/z (ESI): 199&201[ M+1].

Fifth step

6- (4-chlorophenyl) -8, 8-dimethyl-spiro [2.5] oct-5-ene-5-carbaldehyde

6-chloro-8, 8-dimethylspiro [2.5] oct-5-ene-5-carbaldehyde (0.34 g,1.70 mmol), p-chlorophenylboronic acid (0.41 g,2.60 mmol), potassium carbonate (0.70 g,5.10 mmol), tetra-n-butylammonium bromide (0.55 g,1.70 mmol) and palladium acetate (77 mg,0.34 mmol) were added to water (15 mL) under nitrogen. The system is vacuumized and replaced by nitrogen for three times, and the temperature is raised to 50 ℃ for reaction for 4 hours. Cooled to room temperature, the reaction mixture was extracted with ethyl acetate (20 mL. Times.2), the combined organic phases were washed with saturated brine (20 mL), the organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (0-5% ethyl acetate/petroleum ether) to give 6- (4-chlorophenyl) -8, 8-dimethyl spiro [2.5] oct-5-ene-5-carbaldehyde (0.18 g, pale yellow oily substance). Yield: 38.2%. MS m/z (ESI): 275&277[ M+1].

Sixth step

(6- (4-chlorophenyl) -8, 8-dimethyl-spiro [2.5] oct-5-en-5-yl) methanol

6- (4-chlorophenyl) -8, 8-dimethyl-spiro [2.5] oct-5-ene-5-carbaldehyde (0.18 g,0.66 mmol) was dissolved in a tetrahydrofuran-methanol mixed solution (v/v=6/1, 7 ml), and sodium borohydride (75 mg,1.78 mmol) was added thereto and stirred at room temperature for 2 hours. The reaction was quenched by the addition of saturated aqueous ammonium chloride (20 mL), the reaction was extracted with ethyl acetate (20 mL. Times.2), the combined organic phases were washed with saturated brine (20 mL), the organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure to give (6- (4-chlorophenyl) -8, 8-dimethyl spiro [2.5] oct-5-en-5-yl) methanol (0.16 g, colorless oil). Yield: 88.2%. MS m/z (ESI): 260&262[ M-17].

Seventh step

5- (chloromethyl) -6- (4-chlorophenyl) -8, 8-dimethyl spiro [2.5] oct-5-ene

(6- (4-chlorophenyl) -8, 8-dimethyl-spiro [2.5] oct-5-en-5-yl) methanol (0.16 g,0.58 mmol) was dissolved in dichloromethane (10 mL), and thionyl chloride (0.21 g,1.74 mmol) was added and stirred at room temperature overnight. Concentrated under reduced pressure to give 5- (chloromethyl) -6- (4-chlorophenyl) -8, 8-dimethyl spiro [2.5] oct-5-ene (0.17 g, yellow oil). Crude product.

¹ H NMR(400MHz，CDCl ₃ )δ7.36-7.29(m，2H)，7.20-7.13(m，2H)，3.88(s，2H)，2.20-2.11(m，2H)，1.64-1.53(m，2H)，0.85(s，6H)，0.63-0.57(m，2H)，0.24-0.16(m，2H)。

Intermediate 9

7, 8-tetramethyl-1, 4-dioxaspiro [4.5] decane

7, 8-tetramethyl-1, 4-dioxaspiro [4.5] decane

The compound 4, 4-dimethyl-7, 10-dioxadispiro [2.2.4 ⁶ .2 ³ ]Dodecane (see intermediate 8, second step) (4.30 g,21.9 mmol) and platinum dioxide (1.00 g,4.40 mmol) were added to acetic acid (30 mL), and the reaction was purged with hydrogen and then warmed to 40℃for overnight reaction. Cooled to room temperature, diluted with ethyl acetate (30 mL), the reaction mixture was filtered through celite, the filter cake was washed with ethyl acetate (20 mL), and the filtrate was concentrated under reduced pressure to give the crude 7, 8-tetramethyl-1, 4-dioxaspiro [4.5]]Decane (4.50 g, colorless oil). Crude product. MS (MS)m/z(ESI)：199[M+1]。

Synthesis of intermediate 10 refers to the synthesis step of intermediate 8.

Example 1

(R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -3-methylpiperazin-1-yl) -N- ((3-fluoro-5-nitro-4- (((tetrahydro-2H-pyran-4-yl) methyl) amino) phenyl) sulfonyl) benzamide

First step

(R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (3-methylpiperazin-1-yl) benzoic acid methyl ester

Methyl 2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4-fluorobenzoate (synthetic reference: journal of Organic Chemistry,84 (8), 4814-4829; 2019) (2.86 g,10.00 mmol), (R) -2-methylpiperazine (3.00 g,30.00 mmol), N-diisopropylethylamine (3.12 g,24.18 mmol) and dimethyl sulfoxide (20 mL) were mixed. Stirring was carried out at 60℃for 16 hours. The mixture was diluted with 50mL of water and extracted with ethyl acetate (60 mL. Times.2). The combined organic phases were washed with saturated brine (50 mL. Times.3). The organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the crude product was obtained by desolventizing under reduced pressure. Column chromatography purification (dichloromethane/methanol=90:10) afforded the target product, methyl (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (3-methylpiperazin-1-yl) benzoate (2.55 g, yellow solid). Yield: 70%.

MS m/z(ESI)：367[M+1]；

¹ H NMR(400MHz，CDCl ₃ )δ9.92(s，1H)，8.19(d，J＝2.4Hz，1H)，7.91(d，J＝9.2Hz，1H)，7.52(d，J＝2.4Hz，1H)，7.36-7.35(m，1H)，6.66-6.63(m，1H)，6.43(d，J＝1.6Hz，1H)，6.34(d，J＝2.4Hz，1H)，3.79(s，3H)，3.54-3.48(m，2H)，3.05-3.02(m，1H)，2.93-2.84(m，2H)，2.77-2.71(m，1H)，2.42-2.37(m，1H)，1.08(d，J＝6.4Hz，3H)。

Second step

(R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -3-methylpiperazin-1-yl) benzoic acid methyl ester

The compound (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (3-methylpiperazin-1-yl) benzoic acid methyl ester (34 mg,0.09 mmol), 4 '-chloro-6- (chloromethyl) -3, 3-dimethyl-2, 3,4, 5-tetrahydro-1, 1' -biphenyl (synthetic reference: US 20100298323) (30 mg,0.11 mmol), N-diisopropylethylamine (36 mg,0.28 mmol) and acetonitrile (5.0 mL) were mixed. Stirring was carried out at 60℃for 16 hours. The mixture was quenched with 10mL of water and extracted with ethyl acetate (10 mL. Times.2). The combined organic phases were washed with saturated brine (10 mL. Times.2). The organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the crude product was obtained by desolventizing under reduced pressure. Purification with preparative plate (petroleum ether/ethyl acetate=1:1) afforded the target product (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -3-methylpiperazin-1-yl) benzoic acid methyl ester (28 mg, colorless oily liquid). Yield: 50%. MS m/z (ESI): 599&601[ M+1].

Third step

(R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -3-methylpiperazin-1-yl) benzoic acid

The compound (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -3-methylpiperazin-1-yl) benzoic acid methyl ester (28 mg,0.05 mmol), lithium hydroxide (20 mg,0.83 mmol), methanol (1 mL), tetrahydrofuran (1 mL) and water (1 mL) were mixed and stirred at 60℃for 1.5 hours. The mixture was neutralized with 1N hydrochloric acid and extracted with ethyl acetate (10 mL. Times.2). The combined organic phases were washed with saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the crude (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -3-methylpiperazin-1-yl) benzoic acid (25.6 mg, crude) was obtained by desolventizing under reduced pressure. This mixture was used in the next reaction without purification. MS m/z (ESI): 585&587[ M+1].

Fourth step

The compound (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -3-methylpiperazin-1-yl) benzoic acid (26 mg,0.04 mmol), 3-fluoro-5-nitro-4- (((tetrahydro-2H-pyran-4-yl) methyl) amino) benzenesulfonamide (intermediate 1) (15 mg,0.04 mmol), 4-dimethylaminopyridine (11 mg,0.09 mmol), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (17 mg,0.09 mmol) and dichloromethane (2.5 mL) were mixed. Stirring at room temperature for 16 hours. The mixture was quenched with 10mL of water and extracted with dichloromethane (10 mL. Times.2). The combined organic phases were washed with saturated brine (10 mL). The organic phase was dried over anhydrous sodium sulfate, the drying agent was removed by filtration, and the crude product was obtained by desolventizing under reduced pressure. Preparation of silica gel plate purification (dichloromethane/methanol=15:1) afforded the target product (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -3-methylpiperazin-1-yl) -N- ((3-fluoro-5-nitro-4- (((tetrahydro-2H-pyran-4-yl) methyl) amino) phenyl) sulfonyl) benzamide 1 (15.7 mg, yellow solid). Yield: 38%.

MS m/z(ESI)：900&902[M+1]；

¹ H NMR(400MHz，CD ₃ OD)δ8.49(s，1H)，8.08-8.05(m，1H)，7.71-7.64(m，2H)，7.50(s，1H)，7.44(d，J＝3.6Hz，1H)，7.36-7.34(m，2H)，7.09-7.07(m，2H)，6.77(d，J＝8.0Hz，1H)，6.42-6.36(m，2H)，4.04-3.93(m，3H)，3.71-3.67(m，2H)，3.52-3.38(m，5H)，3.28-3.26(m，2H)，3.08-3.02(m，1H)，2.96-2.93(m，1H)，2.65-2.63(m，1H)，2.24-2.15(m，2H)，2.06-2.04(m，1H)，1.93-1.89(m，1H)，1.85(d，J＝4.0Hz，3H)，1.70-1.64(m，3H)，1.58-1.56(m，2H)，1.39-1.28(m，2H)，1.01(s，3H)，0.90(s，3H)。

The synthetic procedure of examples 2 to 7 refers to the procedure of example 1.

Example 8

(R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -2-methylpiperazin-1-yl) -N- ((3-nitro-4- (((tetrahydro-2H-pyran-4-yl) methyl) amino) phenyl) sulfonyl) benzamide

First step

(R) -4- (3- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4-cyanophenyl) -3-methylpiperazine-1-carboxylic acid tert-butyl ester

Compound 2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4-fluorobenzonitrile (synthetic reference: WO 2017212431) (0.51 g,2.00 mmol) and tert-butyl (R) -3-methylpiperazine-1-carboxylate (1.20 g,6.00 mmol) were dissolved in dimethyl sulfoxide (15 mL), dipotassium hydrogen phosphate (1.05 g,6.00 mmol) was added at room temperature and stirred at 110℃for 36 hours. Cooled to room temperature, the reaction mixture was diluted with water, extracted with ethyl acetate (30 ml×2), the combined organic phases were washed with saturated brine (30 ml×2), and the organic phases were dried and concentrated to give the crude product, which was purified by silica gel column chromatography (petroleum ether: ethyl acetate=3:1) to give (R) -4- (3- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4-cyanophenyl) -3-methylpiperazine-1-carboxylic acid tert-butyl ester (0.35 g, white solid) in 41% yield.

MS m/z(ESI)：434[M+1]；

¹ H NMR(400MHz，CDCl ₃ )δ9.29(s，1H)，8.19(s，1H)，7.69(s，1H)，7.49(d，J＝8.8Hz，1H)，7.42-7.40(m，1H)，6.53-6.49(m，2H)，6.08(s，1H)，40.5-3.83(m，3H)，3.21-3.20(m，2H)，3.04-2.95(m，2H)，1.44(s，9H)，1.02(d，J＝6.4Hz，3H)。

Second step

(R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (2-methylpiperazin-1-yl) benzonitrile

(R) -4- (3- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4-cyanophenyl) -3-methylpiperazine-1-carboxylic acid tert-butyl ester (0.35 g,0.81 mmol) was dissolved in dioxane hydrochloride solution (4M, 20.00mmol,5 mL) and stirred at room temperature for 1 hour. The reaction solution was dried by spinning to give the crude product (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (2-methylpiperazin-1-yl) benzonitrile (0.38 g, crude) which was used directly in the next step without purification. MS m/z (ESI): 334[ M+1].

Third step

(R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -2-methylpiperazin-1-yl) benzonitrile

(R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (2-methylpiperazin-1-yl) benzonitrile (0.38 g, crude) and 4 '-chloro-6- (chloromethyl) -3, 3-dimethyl-2, 3,4, 5-tetrahydro-1, 1' -biphenyl (0.21 g,0.80 mmol) (synthetic reference: US 20100298323) were dissolved in acetonitrile (10 mL), N-diisopropylethylamine (0.31 g,2.40 mmol) was added and stirred at 80℃for 4 hours. Cooled to room temperature, the reaction mixture was diluted with water, extracted with ethyl acetate (20 mL. Times.2), and the combined organic phases were washed with saturated brine (20 mL). The organic phase was dried and concentrated to give the crude product, which was purified by silica gel column (petroleum ether: ethyl acetate=2:1) to give (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -2-methylpiperazin-1-yl) benzonitrile (0.30 g, white solid) in 66% yield.

MS m/z(ESI)：566&568[M+1]；

¹ H NMR(400MHz，CDCl ₃ )δ9.40(s，1H)，8.19(s，1H)，7.68(s，1H)，7.43(d，J＝8.8Hz，1H)，7.42-7.40(m，1H)，7.23(d，J＝8.4Hz，2H)，6.92(d，J＝8.4Hz，2H)，6.52-6.45(m，2H)，6.04(s，1H)，3.81-3.78(m，1H)，3.16-3.13(m，1H)，2.98-2.95(m，1H)，2.69-2.65(m，3H)，2.54-2.51(m，1H)，2.21-2.14(m，2H)，2.04-1.97(m，2H)，1.89-1.76(m，2H)，1.43-1.40(m，2H)，1.07(d，J＝6.4Hz，3H)，0.94(s，3H)，0.93(s，3H)。

Fourth step

(R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -2-methylpiperazin-1-yl) benzoic acid

The compound (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -2-methylpiperazin-1-yl) benzonitrile (0.11 g,0.20 mmol) was dissolved in 2-butanol (3 mL) and sodium hydroxide (160 mg,4.00 mmol) was added at 90℃and heated to 105℃with stirring for 24 hours. The reaction mixture was desolventized under reduced pressure, and the residue was taken up in 1N aqueous hydrochloric acid (4 mL) and extracted with ethyl acetate (10 mL. Times.2), and the combined organic phases were washed with saturated brine (10 mL). The organic phase was dried and concentrated to give the crude product, which was purified by prep. silica gel plate (dichloromethane: methanol=10:1) to give (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -2-methylpiperazin-1-yl) benzoic acid (90 mg, white solid) in 77% yield. MS m/z (ESI): 585&587[ M+1].

Fifth step

The final procedure of reference example 1 was synthesized to give the desired product, (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -2-methylpiperazin-1-yl) -N- ((3-nitro-4- (((tetrahydro-2H-pyran-4-yl) methyl) amino) phenyl) sulfonyl) benzamide 8.

MS m/z(ESI)：882&884[M+1]；

¹ H NMR(400MHz，CDCl ₃ )δ10.13(s，1H)，9.29(s，1H)，8.88(s，1H)，8.53-8.50(m，1H)，8.22-8.16(m，2H)，7.94(d，J＝8.8Hz，1H)，7.70(s，1H)，7.47-7.45(m，1H)，7.22(d，J＝8.0Hz，2H)，6.93-6.89(m，3H)，6.56-6.49(m，2H)，5.94(s，1H)，4.05-4.01(m，2H)，3.78-3.74(m，1H)，3.45-3.39(m，2H)，3.28-3.25(m，2H)，3.15-3.12(m，1H)，2.96-2.89(m，1H)，2.73-2.62(m，3H)，2.51-2.48(m，1H)，2.20-2.11(m，2H)，1.99-1.95(m，3H)，1.86-1.83(m，1H)，1.79-1.72(m，3H)，1.45-1.39(m，4H)，1.03(d，J＝6.4Hz，3H)，0.93(s，3H)，0.92(s，3H)。

The synthetic procedure of examples 9 to 13 refers to the procedure of example 8.

The synthetic procedure of examples 14 to 22 refers to the procedure of example 1.

The synthetic procedure of examples 23 to 26 refers to the procedure of example 8.

The synthetic procedure of examples 27 to 30 is referred to in example 1.

Example 31

(R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((6- (4-chlorophenyl) -8, 8-dimethyl-spiro [2.5] oct-5-en-5-yl) methyl) -3-methylpiperazin-1-yl) -N- ((3-nitro-4- (((tetrahydro-2H-pyran-4-yl) methyl) amino) phenyl) sulfonyl) benzamide

First step

(R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((6- (4-chlorophenyl) -8, 8-dimethyl-spiro [2.5] oct-5-en-5-yl) methyl) -3-methylpiperazin-1-yl) benzoic acid methyl ester

Methyl 5- (chloromethyl) -6- (4-chlorophenyl) -8, 8-dimethyl spiro [2.5] oct-5-ene (intermediate 8) (0.17 g,0.58 mmol), (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (3-methylpiperazin-1-yl) benzoate and N, N-diisopropylethylamine (0.37 g,2.90 mmol) were dissolved in acetonitrile (15 mL) and heated to reflux for 3H. After cooling, the mixture was concentrated under reduced pressure, the residue was dissolved in ethyl acetate (30 mL), washed with water (30 mL. Times.3) and saturated brine (30 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel (10% -40% ethyl acetate/petroleum ether) to give methyl (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((6- (4-chlorophenyl) -8, 8-dimethyl spiro [2.5] oct-5-en-5-yl) methyl) -3-methylpiperazin-1-yl) benzoate (0.10 g, white solid). Yield: 27.7%. MS m/z (ESI): 625&627[ M+1].

Second step

(R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((6- (4-chlorophenyl) -8, 8-dimethyl spiro [2.5] oct-5-en-5-yl) methyl) -3-methylpiperazin-1-yl) benzoic acid

Methyl (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((6- (4-chlorophenyl) -8, 8-dimethyl spiro [2.5] oct-5-en-5-yl) methyl) -3-methylpiperazin-1-yl) benzoate (0.10 g,0.16 mmol) was dissolved in ethanol (3 mL) and aqueous sodium hydroxide (2N, 6.00mmol,3 mL) and heated to 70℃for 2 hours. After cooling, the mixture was diluted with water (20 mL), and the pH was adjusted to about 5 by adding 1N diluted hydrochloric acid, and the aqueous phase was extracted with ethyl acetate (20 mL. Times.2). The combined organic phases were washed with saturated brine (20 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate concentrated under reduced pressure to give (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((6- (4-chlorophenyl) -8, 8-dimethyl spiro [2.5] oct-5-en-5-yl) methyl) -3-methylpiperazin-1-yl) benzoic acid (80 mg, white solid). Yield: 81.8%. MS m/z (ESI): 611&613[ M+1].

Third step

The compound (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((6- (4-chlorophenyl) -8, 8-dimethyl-spiro [2.5] oct-5-en-5-yl) methyl) -3-methylpiperazin-1-yl) benzoic acid (20 mg,0.03 mmol), 3-nitro-4- (((tetrahydro-2H-pyran-4-yl) methyl) amino) benzenesulfonamide (synthetic reference: WO 2018041248A 1) (10 mg,0.03 mmol), 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (31 mg,0.17 mmol) and 4-dimethylaminopyridine (8 mg,0.07 mmol) was dissolved in dichloromethane (5 mL) and stirred overnight at room temperature. Dichloromethane (20 mL) was added to dilute the mixture, the reaction mixture was washed with water (20 ml×3) and saturated brine (20 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. Purification of the residue by preparative thin layer chromatography (dichloromethane/methanol=15/1) afforded the target product (R) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- (4- ((6- (4-chlorophenyl) -8, 8-dimethyl spiro [2.5] oct-5-en-5-yl) methyl) -3-methylpiperazin-1-yl) -N- ((3-nitro-4- (((tetrahydro-2H-pyran-4-yl) methyl) amino) phenyl) sulfonyl) benzamide 31 (16.6 mg, yellow solid), yield: 55.7%

MS m/z(ESI)：908&910[M+1]；

¹ H NMR(400MHz，CDCl ₃ )δ10.14(brs，1H)，9.70(s，1H)，8.96-8.82(m，1H)，8.57-8.48(m，1H)，8.27-8.09(m，2H)，8.00-7.90(m，1H)，7.75-7.63(m，1H)，7.50-7.43(m，1H)，7.25-7.19(m，2H)，7.01-6.85(m，3H)，6.59-6.49(m，2H)，5.98(d，J＝2.0Hz，1H)，4.10-3.95(m，2H)，3.48-3.38(m，2H)，3.32-3.17(m，4H)，3.12-3.03(m，1H)，2.87-2.73(m，1H)，2.70-2.45(m，3H)，2.18-2.04(m，3H)，2.03-1.83(m，3H)，1.50-1.19(m，6H)，0.93-0.78(m，8H)，0.60-0.48(m，2H)，0.15-0.02(m，2H)。

The synthetic procedure of examples 32 to 34 refers to the procedure of example 31.

The synthetic procedure of examples 35 to 38 refers to the procedure of example 1.

The synthesis procedure of examples 39 to 42 refers to the procedure of example 31.

The synthetic procedure of examples 43 to 46 refers to the procedure of example 8.

The synthetic procedure of examples 47 to 56 refers to the procedure of example 1.

Example 57

2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- ((4 ar,8 ar) -4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) octahydroquinoxalin-1 (2H) -yl) -N- ((3-nitro-4- (((tetrahydro-2H-pyran-4-yl) methyl) amino) phenyl) sulfonyl) benzamide

Example 57 Synthesis of the Synthesis procedure of reference example 1 in which (4 aR,8 aR) -decahydroquinoxaline (Synthesis reference: european Journal of Organic Chemistry,2012 (34), 6752-6759; 2012) was used in place of (R) -2-methylpiperazine, the second step gave the title compound 2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- ((4 aR,8 aR) -4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) octahydroquinoxalin-1 (2H) -yl) -N- ((3-nitro-4- (((tetrahydro-2H-pyran-4-yl) methyl) amino) phenyl) sulfonyl) benzamide 57.

MS m/z(ESI)：822&824[M+1]；

¹ H NMR(400MHz，CDCl ₃ )δ10.21(s，1H)，8.93-8.92(m，1H)，8.88-8.87(m，1H)，8.57-8.53(m，1H)，8.20-8.18(m，2H)，8.01(d，J＝8.4Hz，1H)，7.71(d，J＝1.6Hz，1H)，7.46-7.44(m，1H)，7.20(d，J＝8.0Hz，2H)，6.93(m，1H)，6.89(d，J＝8.0Hz，2H)，6.73-6.70(m，1H)，6.57-6.56(m，1H)，6.17(d，J＝1.6Hz，1H)，4.06-4.02(m，2H)，3.45-3.41(m，3H)，3.29-3.26(m，2H)，3.19-3.16(m，1H)，3.04-3.01(m，1H)，2.81-2.69(m，2H)，2.49-2.43(m，2H)，2.22-2.19(m，1H)，1.98-1.96(m，4H)，1.96-1.92(m，4H)，1.85-1.82(m，1H)，1.76-1.73(m，3H)，1.47-1.44(m，3H)，1.41-1.35(m，4H)，0.96(s，3H)，0.93(s，3H)。

Example 58

2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- ((2 r,5 r) -4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -2, 5-dimethylpiperazin-1-yl) -N- ((3-nitro-4- (((tetrahydro-2H-pyran-4-yl) methyl) amino) phenyl) sulfonyl) benzamide

EXAMPLE 58 Synthesis of reference example 1 the procedure of example 1 was followed, in which (2R, 5R) -2, 5-dimethylpiperazine (synthetic reference: organic Chemistry Frontiers,5 (23), 3402-3405; 2018) was used instead of (R) -2-methylpiperazine, to give the title compound 2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- ((2R, 5R) -4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -2, 5-dimethylpiperazin-1-yl) -N- ((3-nitro-4- (((tetrahydro-2H-pyran-4-yl) methyl) amino) phenyl) sulfonyl) benzamide 58.

MS m/z(ESI)：896&898[M+1]；

¹ H NMR(400MHz，CDCl ₃ )δ10.10(s，1H)，9.25(s，1H)，8.88(d，J＝2.2Hz，1H)，8.53-8.51(m，1H)，8.20(d，J＝2.5Hz，1H)，8.19-8.02(m，1H)，7.94(d，J＝9.2Hz，1H)，7.70(d，J＝2.4Hz，1H)，7.48-7.42(m，1H)，7.22(d，J＝8.4Hz，2H)，6.99-6.87(m，2H)，6.59-6.52(m，1H)，6.50-6.40(m，1H)，5.95(d，J＝2.2Hz，1H)，4.10-4.01(m，2H)，3.70-3.60(m，2H)，3.47-3.37(m，2H)，3.33-3.21(m，2H)，3.19-3.03(m，2H)，2.59-2.50(m，2H)，2.39-2.31(m，2H)，1.97-1.90(m，3H)，1.82-1.71(m，2H)，1.46-1.33(m，3H)，1.28-1.20(m，3H)，1.25-1.15(m，4H)，1.02-1.00(m，2H)，0.95(s，3H)，0.93(s，3H)。

Example 59

N- ((4- ((((S) -1, 4-dioxan-2-yl) methyl) amino) -3-nitrophenyl) sulfonyl) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- ((3R, 5R) -4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -3, 5-dimethylpiperazin-1-yl) benzamide

Example 59 Synthesis of reference example 1 the procedure of example 1 was followed, in which (2R, 6R) -2, 6-dimethylpiperazine was used in place of (R) -2-methylpiperazine, to give the target compound N- ((4- ((((S) -1, 4-dioxan-2-yl) methyl) amino) -3-nitrophenyl) sulfonyl) -2- ((1H-pyrrolo [2,3-b ] pyridin-5-yl) oxo) -4- ((3R, 5R) -4- ((4 '-chloro-5, 5-dimethyl-3, 4,5, 6-tetrahydro- [1,1' -biphenyl ] -2-yl) methyl) -3, 5-dimethylpiperazin-1-yl) benzamide 59.

MS m/z(ESI)：898&890[M+1]；

¹ H NMR(400MHz，CDCl ₃ )δ9.16(s，1H)，8.86(d，J＝2.2Hz，1H)，8.61(s，1H)，8.22-8.13(m，2H)，7.91(d，J＝9.1Hz，1H)，7.67-7.60(m，1H)，7.44(s，1H)，7.23(d，J＝8.3Hz，2H)，6.93(d，J＝8.3Hz，2H)，6.88(d，J＝9.2Hz，1H)，6.60-6.48(m，2H)，5.94-5.90(m，1H)，3.97-3.85(m，2H)，3.85-3.74(m，2H)，3.67-3.60(m，2H)，3.55-3.29(m，3H)，3.13-3.02(m，2H)，2.79-2.70(m，2H)，2.69-2.59(m，2H)，2.24-2.15(m，2H)，2.12-2.05(m，4H)，2.05-1.86(m，2H)，0.93-0.91(m，6H)，0.69(s，3H)，0.67(s，3H)。

Biological experiments

Test for inhibition of biological activity of BCL-2

The experimental methods for assessing the effect of the compounds of the invention on BCL-2 bioactivity using fluorescence polarization experiments are summarized below:

the effect of the compounds on the biological activity of BCL-2 was evaluated by detecting the effect of the compounds on the binding activity of BCL-2 and leukemia pro-apoptotic proteins (BIMs) using an affinity assay method based on fluorescence polarization principle. The reaction buffer contained the following components: PBS (pH 7.4,3mM Na) ₂ HPO ₄ 、155mM NaCl、1mM KH ₂ PO ₄ ) 1mM DTT; human recombinant Bcl-2 protein (cat No. 10195-H08E) was purchased from Beijing Yiqiao Shenzhou biotechnology Co., ltd, diluted to 5nM with reaction buffer; FITC-labeled BIM polypeptides were purchased from Nanjing gold St Biotechnology Co., ltd and diluted to 5nM with reaction buffer.

Compounds were dissolved and diluted to 0.1, 1, 10. Mu.M in 100% DMSO, then serially diluted 4-fold with DMSO to a minimum concentration of 0.0061, 0.061, 0.61nM, and diluted 50-fold with reaction buffer at each concentration point.

To a black 384-well assay plate, 3 μl of compound solution and 12 μl of BCL-2 solution were added, mixed well and incubated at room temperature for 15 minutes. Subsequently, 15. Mu.L of FITC-BIM solution was added, and the reaction mixture was incubated at room temperature for 30 minutes in the absence of light, immediately followed by fluorescence polarization detection on an Envision multifunctional enzyme-labeled instrument (Perkin Elmer), 480nm being the excitation wavelength and 535nm being the emission wavelength. In this experiment, the group without BCL-2 protein was used as a negative control (100% inhibition), and the group with BCL-2 protein but without compound was used as a positive control (0% inhibition). The percent inhibition of BCL-2 affinity by a compound can be calculated by the following formula:

Compound IC50 values were calculated from 8 concentration points using XLfit (ID Business Solutions ltd., UK) software by the following formula:

Y＝Bottom+(Top-Bottom)/(1+10^((logIC ₅₀ -X)*slope factor))

wherein Y is the inhibition percentage, X is the logarithmic value of the concentration of the compound to be detected, bottom is the maximum inhibition percentage, top is the minimum inhibition percentage, and slope factor is the curve slope coefficient.

Test for inhibition of biological activity of BCL-XL

Evaluation of the Effect of the inventive Compounds on the biological Activity of BCL-XL Using fluorescence polarization experiments

The experimental procedure is summarized as follows:

the effect of the compounds on BCL-XL biological activity was assessed by detecting the effect of the compounds on BCL-XL and BIM binding activity using an affinity assay method using fluorescence polarization principle. The reaction buffer contained the following components: PBS (pH 7.4,3mM Na) ₂ HPO ₄ 、155mM NaCl、1mM KH ₂ PO ₄ ) 1mM DTT; human recombinant Bcl-XL protein (cat. No. 10455-H08E) was purchased from Beijing Yiqiao Shenzhou biotechnology Co., ltd, diluted to 10nM with reaction buffer; FITC-labeled BIM polypeptides were purchased from Nanjing gold St Biotechnology Inc., and diluted to 10nM with reaction buffer.

Compounds were diluted to 1 μm in 100% DMSO, then serially diluted 4-fold with DMSO to a minimum concentration of 0.061nM, and diluted 50-fold with reaction buffer at each concentration point.

To a black 384-well assay plate, 3 μl of compound solution and 12 μl of BCL-XL solution were added, mixed well and incubated at room temperature for 15 minutes. Subsequently, 15. Mu.L of FITC-BIM solution was added, and the reaction mixture was incubated at room temperature for 30 minutes in the absence of light, immediately followed by fluorescence polarization detection on an Envision multifunctional enzyme-labeled instrument (Perkin Elmer), 480nm being the excitation wavelength and 535nm being the emission wavelength. In this experiment, the group without BCL-XL protein was used as negative control (100% inhibition), the group with BCL-XL protein but without compound was used as positive control (0% inhibition). The percent inhibition of BCL-XL affinity by a compound can be calculated by the following formula:

compound IC ₅₀ The values were calculated from 8 concentration points using XLfit (ID Business Solutions ltd., UK) software by the following formula:

Y＝Bottom+(Top-Bottom)/(1+10^((logIC ₅₀ -X)*slope factor))

RS4; concentration of half-effective inhibition of 11 cells (acute lymphoblastic leukemia cells) IC ₅₀ Is (are) determined by

Evaluating the compound pair RS4 of the present invention using a luminescence cell viability assay; 11 effects of cell proliferation.

The experimental procedure is summarized as follows:

Using CellTilter-Glo (CTG) detection kit, detecting the key index ATP of the metabolism of living cells by adopting a unique, high-sensitivity and stable luciferase, wherein the luminous signal generated in the test is in direct proportion to the number of the living cells in the culture medium, so as to detect RS4;11, and a cell proliferation status.

The CellTilter-Glo reagent (Promega, G7572) consists of CellTilter-Glo freeze-dried powder and CellTilter-Glo buffer, and the freeze-dried powder is dissolved into the buffer when in use.

RS4;11 cells (ATCC, CRL-1873) were cultured in RPMI1640 complete medium (Thermofiser, 72400-047) containing 10% FBS (GBICO, 10099-141) and 100units/ml of a mixture of Streptomyces lividans (Thermofiser, 15140122), when the coverage of the cells in the culture vessel reached 80-90%, digested with 0.25% pancreatin (EDTA) (Thermofiser, 25200056) and then blow-dispersed and planted in a white 384-well plate (Thermofiser, 164610), and then the 384-well plate was placed at 37℃with 5% CO ₂ Is cultured overnight in an incubator. Compounds were dissolved and diluted to 5mM in 100% DMSO, then serially diluted 4-fold with DMSO to a minimum concentration of 0.061. Mu.M, and each concentration point was re-used with FBS-free RPMI1640 medium was diluted 50-fold. If compound IC ₅₀ The values are very low and the initial concentration of the compound can be reduced. After overnight, 3 μl of the diluted compound per well was added, and gently centrifuged and mixed, wherein no cell group was added as negative control (100% inhibition) and 0.2% dmso group was added as positive control (0% inhibition). The 384-well plate was placed at 37℃with 5% CO ₂ After 48 hours, the incubator was equilibrated to room temperature, 15. Mu.l of CTG reagent was added to each well, and the mixture was gently shaken on a shaker for 3 minutes to ensure sufficient cell lysis, allowed to stand for 10 minutes to stabilize the luminescence signal, and then the luminescence signal was read by EnVision (Perkin Elmer).

Compound pair RS4; the percentage of 11 cell proliferation inhibition can be calculated by the following formula:

percentage of inhibition = 100-100 x (signal compound-signal negative control)/(signal positive control-signal negative control)

Y＝Bottom+(Top-Bottom)/(1+10^((LogIC ₅₀ -X)*slope factor))

where Y is the percent inhibition, bottom is the Bottom plateau of the curve (Bottom plateau of the S-curve), top is the Top plateau of the curve (Top plateau of the S-curve), and X is the log of the concentration of the test compound.

The results of the in vitro BCL-2 and BCL-XL protein activity assay described above are shown in Table 1 below,

The results of the cell experiments are shown in Table 2.

Table 1: results of detection of the activity of the BCL-2 and BCL-XL proteins

Numbering of compounds	FP BCL-2IC ₅₀ (nM)	FP BCL-XL IC ₅₀ (nM)
1	2.05	＞1000
2	2.56	＞1000
3	3.06	＞1000
4	2.29	742.8
5	1.54	＞1000
6	0.90	＞1000
7	＞100	＞1000
8	4.55	＞1000
9	5.03	＞1000
10	8.21	＞1000
11	12.8	＞1000
12	4.89	＞1000
13	＞100	＞1000
14	2.35	＞1000
15	5.99	＞1000

16	3.41	＞1000
17	1.46	952.96
18	1.32	＞1000
19	2.13	＞1000
20	2.57	＞1000
21	1.97	＞1000
22	1.49	＞1000
23	9.37	＞1000
24	6.3	＞1000
25	2.71	＞1000
26	1.63	＞1000
27	5.12	＞1000
28	1.37	＞1000
29	1.6	＞1000
30	0.84	830.51
31	3.57	＞1000
32	1.8	＞1000
33	3.16	＞1000
34	3.33	＞1000
35	2.96	＞1000
36	4.78	＞1000
37	3.15	＞1000
38	5.24	＞1000
39	2.68	＞1000
40	1.77	＞1000
41	1.28	＞1000
42	1.53	＞1000
43	6.33	＞1000
44	4.98	＞1000
45	4.22	＞1000
46	1.09	＞1000

47	＞100	＞1000
48	63.79	＞1000
49	＞100	＞1000
50	＞100	＞1000
51	＞100	＞1000
52	1.9	＞1000
53	＞100	＞1000
54	11	＞1000
55	2.3	＞1000
56	82	＞1000
57	＞500	＞1000
58	29.86	＞1000
59	73.22	＞1000

。

Table 2: RS4;11 cell Activity assay results

Numbering of compounds	RS4；11IC ₅₀ (nM)
1	4.73
2	11.07
3	6.4
4	5.8
5	2.75
6	2.37
7	＞500
8	6.2
9	26
10	68
11	42
12	16
13	＞500
14	3.8
15	19

16	8.8
17	3.1
18	2.4
19	4.25
20	8.8
21	8.8
22	4.1
23	51
24	24
25	32
26	14
27	16
28	6.55
29	4.2
30	2.3
31	8.1
32	6.6
33	16
34	6.6
35	17
36	21
37	22
38	32
39	14.5
40	15
41	8
42	10
43	161
44	54
45	52
46	18

47	486
48	229
49	＞500
50	＞500
51	＞500
52	5.1
53	＞500
54	25
55	5.7
56	224
57	＞1000
58	141.94
59	193
Venetoclax	5.5

。

From the experimental results, the compounds of the embodiment of the invention can effectively and selectively inhibit the activity of BCL-2, and have weak inhibition on BCL-XL. Can be used for treating various cancers caused by abnormal overexpression of BCL-2 family proteins: especially acute lymphoblastic leukemia, such as hematopathy, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer, and brain glioma. And avoids toxic side effects caused by BCL-XL inhibition, such as thrombocytopenia. Some compounds are also effective in inhibiting RS4;11 acute lymphocyte proliferation. Has strong inhibiting effect on malignant hematopathy such as acute lymphoblastic leukemia, etc.

It will be apparent to those skilled in the art that the present disclosure is not limited to the illustrative embodiments described above, and that it may be embodied in other specific forms without departing from the essential characteristics thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, reference being made to the appended claims rather than to the foregoing embodiments, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.

Claims

A compound of formula (I), an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof,

wherein:

X ¹ a 6 membered saturated heterocyclic group selected from optionally substituted containing one or two heteroatoms selected from N, O, S, wherein the optional substituent is selected from 4 membered saturated heterocyclic groups containing one or two heteroatoms selected from N, O, S; preferably X ¹ A 6 membered saturated heterocyclic group selected from optionally substituted containing one or two heteroatoms selected from N, O, wherein the optional substituent is selected from oxetanyl; further preferably, X ¹ Selected from 1, 4-dioxanyl, tetrahydropyranyl, N-oxetanyl piperidinyl, N-oxetanyl morpholinyl; most preferably, X ¹ Selected from (S) -1, 4-dioxane-2-yl, (R) -1, 4-dioxane-2-yl, tetrahydropyran-4-yl, 1- (oxetan-3-yl) piperidin-4-yl, (S) -4- (oxetan-3-yl) morpholin-2-yl;

X ² a 5-6 membered heterocyclic group selected from the group consisting of containing one or two N atoms; wherein the ring is optionally substituted with one or two C1-C4 alkyl groups; preferably X ² A 6 membered heterocyclic group selected from one or two N atoms; wherein the ring is optionally substituted with one or two C1-C4 alkyl groups; further preferably, X ² Selected from the group consisting of
Wherein the ring is optionally substituted with a C1-C4 alkyl group; most preferably, X ² Selected from the group consisting of

R ⁰ Selected from hydrogen, halogen; preferably, R ⁰ Selected from hydrogen, fluorine, chlorine; most preferably, R ⁰ Selected from hydrogen, fluorine;

R ¹ 、R ² 、R ³ 、R ⁴ each independently selected from hydrogen, C1-C6 alkyl, wherein R ¹ And R is ² Or R is ³ And R is ⁴ Can form 3-6 membered cycloalkyl together with the attached carbon atom; preferably, R ¹ 、R ² 、R ³ 、R ⁴ Each independently selected from hydrogen, C1-C4 alkyl, wherein R ¹ And R is ² Or R is ³ And R is ⁴ Can form 3-4 membered cycloalkyl together with the attached carbon atom; further preferably, R ¹ 、R ² 、R ³ 、R ⁴ Each independently selected from hydrogen or methyl and R ¹ 、R ² 、R ³ 、R ⁴ Not all hydrogen, wherein R ¹ And R is ² Or R is ³ And R is ⁴ Can form a cyclopropyl group together with the attached carbon atom; most preferably, R ¹ 、R ² 、R ³ 、R ⁴ Each independently selected from hydrogen or methyl and R ¹ 、R ² 、R ³ 、R ⁴ Not all hydrogen, wherein R ³ And R is ⁴ Cyclopropyl may be formed together with the attached carbon atom.
A compound of formula (I) according to claim 1, an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, wherein:

X ¹ a 6 membered saturated heterocyclic group selected from optionally substituted containing one or two heteroatoms selected from N, O, wherein the optional substituent is selected from oxetanyl;

X ² A 6 membered heterocyclic group selected from one or two N atoms; wherein the ring is optionally substituted with one or two C1-C4 alkyl groups;

R ⁰ selected from hydrogen, halogen;

R ¹ 、R ² 、R ³ 、R ⁴ each independently selected from hydrogen, C1-C4 alkyl, wherein R ¹ And R is ² Or R is ³ And R is ⁴ Can together with the attached carbon atom form a 3-to 4-membered cycloalkyl group.
A compound of formula (I) according to claim 1, an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, wherein:

X ¹ a 6 membered saturated heterocyclic group selected from optionally substituted containing one or two heteroatoms selected from N, O, wherein the optional substituent is selected from oxetanyl;

X ² is that
Wherein the ring is optionally substituted with a C1-C4 alkyl group;

R ⁰ selected from hydrogen, fluorine, chlorine;

R ¹ 、R ² 、R ³ 、R ⁴ each independently selected from hydrogen, C1-C4 alkyl, wherein R ¹ And R is ² Or R is ³ And R is ⁴ Can together with the attached carbon atom form a 3-to 4-membered cycloalkyl group.
A compound of formula (I) according to claim 1, an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, wherein:

X ¹ selected from 1, 4-dioxanyl, tetrahydropyranyl, N-oxetanyl piperidinyl, N-oxetanyl morpholinyl;

X ² Is that
Wherein the ring is optionally substituted with a C1-C4 alkyl group;

R ⁰ selected from hydrogen, fluorine, chlorine;

R ¹ 、R ² 、R ³ 、R ⁴ each independently selected from hydrogen or methyl and R ¹ 、R ² 、R ³ 、R ⁴ Not all hydrogen, wherein R ¹ And R is ² Or R is ³ And R is ⁴ Cyclopropyl may be formed together with the attached carbon atom.
A compound of formula (I) according to claim 1, an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, wherein:

X ¹ selected from 1, 4-dioxanyl, tetrahydropyranyl, N-oxetanyl piperidinyl, N-oxetanyl morpholinyl;

X ² selected from the group consisting of

R ⁰ Selected from hydrogen, fluorine;

R ¹ 、R ² 、R ³ 、R ⁴ each independently selected from hydrogen or methyl and R ¹ 、R ² 、R ³ 、R ⁴ Not all hydrogen, wherein R ¹ And R is ² Or R is ³ And R is ⁴ Cyclopropyl may be formed together with the attached carbon atom.
A compound of formula (I) according to claim 1, an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, wherein:

X ¹ selected from (S) -1, 4-dioxane-2-yl, (R) -1, 4-dioxane-2-yl, tetrahydropyran-4-yl, 1- (oxetan-3-yl) piperidin-4-yl, (S) -4- (oxetan-3-yl) morpholin-2-yl;

X ² selected from the group consisting of

R ⁰ Selected from hydrogen, fluorine;

R ¹ 、R ² 、R ³ 、R ⁴ each independently selected from hydrogen or methyl and R ¹ 、R ² 、R ³ 、R ⁴ Not all hydrogen, wherein R ³ And R is ⁴ Cyclopropyl may be formed together with the attached carbon atom.
A compound of formula (I) according to claim 1, an isomer, prodrug, solvate, stable isotope derivative or a pharmaceutically acceptable salt thereof, selected from:
use of a compound according to any one of claims 1-7, or an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, in the manufacture of a medicament for use as a BCL-2 inhibitor.
Use of a compound according to any one of claims 1-7, or an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment or prophylaxis of a disease associated with BCL-2, such as a tumor, for example, hematological malignancy of acute lymphoblastic leukemia, lung cancer, breast cancer, ovarian cancer, rectal cancer, prostate cancer, pancreatic cancer, brain glioma.
A pharmaceutical composition comprising a compound according to any one of claims 1-7 or an isomer, prodrug, solvate, stable isotope derivative or pharmaceutically acceptable salt thereof, optionally one or more other BCL-2 inhibitors, and one or more pharmaceutically acceptable carriers, diluents and excipients.